AZA TREE KANGAROO SSP© POST MORTEM PROTOCOL

The following pages contain an SSP approved tree kangaroo post mortem protocol. It is imperative that all tree kangaroos be necropsied, including neonatal carcasses (See below).

Please fill out the appropriate forms. It is recommended that copies of this data be filed with the Animal Health Department at your institution. Send a copy of the final pathology report to Jacque Blessington, Tree Kangaroo SSP© Coordinator, Kansas City Zoological Gardens, 6700 Zoo Drive, Kansas City, MO 64132, Phone: 816-513-5700, FAX 816-513-4654, email jacsprat65@aol.com.

Post mortem radiographs are recommended, whenever possible, to document bony and/or soft tissue lesions (i.e. arthritis).

(The addendums referred to in these pages are from the 2007 Tree Kangaroo Husbandry Manual)

D4

ADDENDUM 4.4: Tree Kangaroo SSP© Necropsy Protocol

Dear Colleague;

There is continued concern about Mycobacterium avium in tree kangaroos, because of an increased awareness of the condition associated with tree kangaroo deaths. It is well to remember that M. avium has been a historical problem in this species and other marsupials, and that the organism is wide spread in the environment.

M. avium infections in tree kangaroos usually present with signs of weight loss (be sure to check for dental disease) plus occasional coughing. Pulmonary lesions have been seen radiographically but are not pathognomonic. At this time we are uncertain if finding M. avium in a tracheal wash indicates infections or colonization of the organism! A good premise to follow is that with a sick tree kangaroo one should have M. avium high on the list as a differential diagnosis until proved otherwise.

At the previous tree kangaroo SSP meeting there was discussion on developing a uniform screening protocol for animals scheduled for shipment and as part of a routine physical. The problem is we do not know yet what tests and/or procedures will allow us to detect infected individuals. For this reason we think it may be premature to establish a rigid protocol at this time. Our work with this disease in tree kangaroos, in collaboration with several investigators in the Washington D.C. medical community, is providing us with several new and exciting approaches to the problems we currently face with this disease. This includes methods of diagnosis, treatment and prevention.

It is recommended that instead of developing a protocol or regulations at this point that we discuss each case individually since any approach we use at this time will be in a state of flux as a result of the new information we are developing. It would also be important to coordinate our efforts through one or two investigators so we can gather significant data as rapidly as possible. We therefore would encourage direct contact with either of us to discuss specific cases.

A necropsy of all animals is essential with collection of a complete set of tissues for histopathological study, plus the appropriate culturing and freezing (-70 C) of selected tissues especially when M. avium is suspected as per the Tree Kangaroo SSP pathology protocol.

If you have any further suggestions or comments on this approach please do not hesitate to contact us. Thank You!

Sincerely;

Mitchell Bush, DVM email giraffedoc@hotmail.com Darin Collins, DVM email darin.collins@zoo.org

ADDENDUM 4.5: Tree Kangaroo SSP© Necropsy Protocol

INSTITUTION/OWNER___________________________________________________

CONTACT PERSON __________________________________________________________ __________

TEL NO. FAX NO._____________________.

EMAIL__________________________

ADDRESS______________________________________________________________

STATE____________________ COUNTRY___________________________________

SPECIES________________________________________________________________

ID# ____________ TRANSPONDER# ______________________ ISIS# ____________ STUDBOOK# ____________ BIRTH DATE ____________ SEX ____________ WEIGHT (kg)____________ (actual/estimate)(circle one)

DEATH DATE___________ DEATH LOCATION ___________________________________

NECROPSY DATE____________ NECROPSY LOCATION ____________________ POST MORTEM INTERVAL ____________

CAPTIVE BORN/LOCATION? ___________________________________________ WILD CAUGHT? ____________

HISTORY(Include clinical signs, circumstances of death, clinical labwork, diet & housing, recent treatments, cagemates, mixed species exhibit, temperature./humidity)

GROSS EXAMINATION(if no abnormalities are noted, mark as normal or NE if not examined)

GENERAL EXAM (Physical and nutritional condition, pelage, subcutaneous fat stores, body orifices, superficial lymph nodes)

MUSCULOSKELETAL SYSTEM (Bones, marrow, joints, muscle)

BODY CAVITIES (fat stores, pleura, thymus, lymph nodes)SPLEEN

RESPIRATORY SYSTEM (nasal passages, pharynx, larynx, trachea, bronchi, lungs, regional lymph nodes)

CARDIOVASCULAR SYSTEM (heart, pericardial sac, great vessels, myocardium, valves, chambers)

DIGESTIVE SYSTEM (mouth, teeth, tongue, esophagus, stomach, small and large intestine, anus, liver and gall bladder, pancreas, mesenteric lymph nodes)

URINARY SYSTEM (kidneys, ureters, bladder, urethra)

REPRODUCTIVE SYSTEM (testes/ovaries, uterus & cervix, penis/vagina, accessory sex organs, mammary gland, placenta)

ENDOCRINE SYSTEM (thyroids, parathyroids, adrenals, pituitary)

CENTRAL NERVOUS SYSTEM (brain, meninges, spinal cord)

SENSORY ORGANS (eyes, ears)

ADDITIONAL COMMENTS OR OBSERVATIONS:

Prosector: Date: ____ ___

SUMMARIZE PRELIMINARY DIAGNOSES:

LABORATORY STUDIES: Results of cytology, fluid analysis, urinalysis, serum chemistries, bacteriology (especially Mycobacterium), mycology, virology, parasitology (ecto and endo parasites), x-ray, photography, other ___________________

ADDENDUM 4.6: TREE KANGAROO SSP© NECROPSY REPORT

TISSUE CHECK LIST

Where possible freeze 3-5 cm block of tissue from major organs (e.g., lung, liver, kidney, spleen) in small plastic bags, preferably in liquid nitrogen to be kept ultrafrozen at -70 degrees Celsius; freeze at conventional temperatures if there is no access to an ultrafreezer.

Preserve as many of the following tissues as possible in 10% buffered formalin at a ratio of approximately 1 part tissue to 10 parts solution. Tissues should be no thicker than .5 to 1 cm. NOTE: There is generally no need to fix and label each tissue separately. Also, freeze post-mortem serum (from heart), urine and any abnormal fluid accumulations for your Pathologist. Consult the Veterinary Advisory Group for any special protocols and instructions about specimens requested by the designated researcher.

Brain Diaphragm Testes/OvariesNerve (Sciatic) Liver UterusSpinal Cord Gall Bladder Mammary GlandEye Spleen UreterTongue Pancreas Urinary BladderEsophagus Stomach UrethraTrachea Small Intestine KidneyThyroid Large Intestine AdrenalParathyroid Cecum ThymusPituitary Skin (axillary) ProstateHeart Aorta Lymph NodeMuscle (pectoral) Bone with marrow Salivary GlandLungs (sternum and rib)

Primary Pathologist: Name Lab Address

Phone

Please attach final pathology report and send a copy with this protocol to Jacque Blessington, Tree Kangaroo SSP© Coordinator, Kansas City Zoo, 6800 Zoo Drive, Kansas City, MO 64132, Phone: 816-513-5800, FAX 816-513-4654, email jacsprat65@aol.com.

ADDENDUM 4.7: Guidelines for Tree Kangaroo SSP© Neonatal Necropsies

The following list includes additional information that should be emphasized and obtained from neonatal animals (including aborted fetuses, stillbirth and neonates up to age of permanent pouch exit). Examine all specimens submitted including partially consumed carcasses. Use this in conjunction with the Tree Kangaroo SSP Necropsy Protocol (Addendum 4.5).

1. Obtain weight, sex; estimate degree of maturity/ immaturity.

2. Crown-rump, tail, hind-leg measurements.

3. Examine skin; pelage - texture, color and amount (if any) of fur; nails - pigmented or not.

4. Examine for external malformations (check for cleft palate, hare lip, other facial/skull, trunk or limb abnormalities.

5. Assess state of hydration (subcutaneous and serosal surfaces dry or moist) and nutritional status (record subcutaneous and body cavity fat stores as none, minimal, moderate, abundant).

6. Examine for internal malformations (e.g. diaphragmatic hernia, cardiac anomalies, etc.).

7. Determine if breathing occurred using the “sink test” by placing a segment of lung in buffered formalin: if lung floats, animal probably breathed. If lung sinks, animal probably did not breathe (if lung proves not to be pneumonic).

8. Verify sex by examining gonads.

9. Determine nursing activity by looking for and estimating amount of milk curd (white, cottage-cheese like) present in stomach and presence of milk stool (yellow-white semisolid material in colon) with absence of meconium (greenish-brown pasty material throughout GI tract).

10. Document degree of trauma induced by dam or other cagemates.

11. Proceed as with standard Tree Kangaroo SSP Necropsy Protocol. Be sure to fix any “placental/ membrane material” if available.

Send a copy of the necropsy report to:

Jacque Blessington, Tree Kangaroo SSP© Coordinator, Kansas City Zoo, 6800 Zoo Drive, Kansas City, MO 64132, Phone: 816-513-5800, FAX 816-513-4654, email jacsprat65@aol.com

TAXIDERMY

The first priority of the TKSSP is that all tree kangaroos be necropsied. After necropsy, the TKSSP requests that the remains of the animal be sent for taxidermy. The following information was received from the Burke Museum.

The Burke Museum at the University of Washington, in Seattle, is very interested in obtaining the tree kangaroo carcasses from AZA institutions. They will prepare them and then make them available for any researchers. Following is information from Jeff Bradley at the Burke Museum, University of Washington, Seattle, WA: We would be happy to receive any tree kangaroos you could send to us; the material (tissue, skeleton, and skin if we made a skin) would be available to researchers after it is accessioned into the Burke collection.

Of primary importance is that the carcasses are frozen as quickly as possible after death, and whatever thawing that happens (e.g., for necropsy) be minimized. For shipping, these specimens should be sent in a cooler with dry ice, overnight delivery, notifying me before shipment so I can be looking for them to arrive. It is best to have them arrive on a Tue, Wed or Thurs (not Friday, because on Fridays small problems with the carrier turn into a 3-day delay instead of just a 1-day delay). See below for my package-shipping address.

Also, if the damage to the skull and skin are minimized during necropsy, that also increases the value of the final specimens. Sometimes skins are cut so much during necropsy that we cannot make a study skin from the specimen, and also big dramatic saw-cuts in the cranium make a specimen less appropriate for showing school children. We understand that necropsies are important - but whatever effort can be made to limit visible damage is valuable to the final museum specimen.

Shipping Address for package deliveries:Jeffrey E. BradleyMammalogy Collection ManagerBurke Museum MammalogyUniv. of WashingtonBox 35301017th Ave NE at NE 45th StSeattle, WA 98195-3010Phone: 206.685.7417 fax: 206.685.3039